A variety of disorders in humans and other mammals involve or are associated with abnormal bone resorption. Such disorders include, but are not limited to, osteoporosis, glucocorticoid induced osteoporosis, Paget""s disease, abnormally increased bone turnover, periodontal disease, tooth loss, bone fractures, rheumatoid arthritis, osteoarthritis, periprosthetic osteolysis, osteogenesis imperfecta, metastatic bone disease, hypercalcemia of malignancy, and multiple myeloma. One of the most common of these disorders is osteoporosis, which in its most frequent manifestation occurs in postmenopausal women. Osteoporosis is a systemic skeletal disease characterized by a low bone mass and microarchitectural deterioration of bone tissue, with a consequent increase in bone fragility and susceptibility to fracture. Osteoporotic fractures are a major cause of morbidity and mortality in the elderly population. As many as 50% of women and a third of men will experience an osteoporotic fracture. A large segment of the older population already has low bone density and a high risk of fractures. There is a significant need to both prevent and treat osteoporosis and other conditions associated with bone resorption. Because osteoporosis, as well as other disorders associated with bone loss, are generally chronic conditions, it is believed that appropriate therapy will typically require chronic treatment.
Osteoporosis is characterized by progressive loss of bone architecture and mineralization leading to the loss in bone strength and an increased fracture rate. The skeleton is constantly being remodeled by a balance between osteoblasts that lay down new bone and osteoclasts that breakdown, or resorb, bone. In some disease conditions and advancing age the balance between bone formation and resorption is disrupted; bone is removed at a faster rate. Such a prolonged imbalance of resorption over formation leads to weaker bone structure and a higher risk of fractures.
Bone resorption is primarily performed by osteoclasts, which are multinuclear giant cells. Osteoclasts resorb bone by forming an initial cellular attachment to bone tissue, followed by the formation of an extracellular compartment or lacunae. The lacunae are maintained at a low pH by a proton-ATP pump. The acidified environment in the lacunae allows for initial demineralization of bone followed by the degradation of bone proteins or collagen by proteases such as cysteine proteases. See Delaisse, J. M. et al., 1980, Biochem J 192:365-368; Delaisse, J. et al., 1984, Biochem Biophys Res Commun:441-447; Delaisse, J. M. et al., 1987, Bone 8:305-313, which are hereby incorporated by reference in their entirety. Collagen constitutes 95% of the organic matrix of bone. Therefore, proteases involved in collagen degradation are an essential component of bone turnover, and as a consequence, the development and progression of osteoporosis.
Cathepsins belong to the papain superfamily of cysteine proteases. These proteases function in the normal physiological as well as pathological degradation of connective tissue. Cathepsins play a major role in intracellular protein degradation and turnover and remodeling. To date, a number of cathepsin have been identified and sequenced from a number of sources. These cathepsins are naturally found in a wide variety of tissues. For example, cathepsin B, F, H, L, K, S, W, and Z have been cloned. Cathepsin K (which is also known by the abbreviation cat K) is also known as cathepsin O and cathepsin O2. See PCT Application WO 96/13523, Khepri Pharmaceuticals, Inc., published May 9, 1996, which is hereby incorporated by reference in its entirety. Cathepsin L is implicated in normal lysosomal proteolysis as well as several disease states, including, but not limited to, metastasis of melanomas. Cathepsin S is implicated in Alzheimer""s disease and certain autoimmune disorders, including, but not limited to juvenile onset diabetes, multiple sclerosis, pemphigus vulgaris, Graves"" disease, myasthenia gravis, systemic lupus erythemotasus, rheumatoid arthritis and Hashimoto""s thyroiditis; allergic disorders, including, but not limited to asthma; and allogenic immunbe responses, including, but not limited to, rejection of organ transplants or tissue grafts.
Cysteine protease inhibitors such as E-64 (trans-epoxysuccinyl-L-leucylamide-(4-guanidino) butane) are known to be effective in inhibiting bone resorption. See Delaisse, J. M. et al., 1987, Bone 8:305-313, which is hereby incorporated by reference in its entirety. Recently, cathepsin K was cloned and found specifically expressed in osteoclasts See Tezuka, K. et al., 1994, J Biol Chem 269:1106-1109; Shi, G. P. et al., 1995, FEBS Lett 357:129-134; Bromme, D. and Okamoto, K., 1995, Biol Chem Hoppe Seyler 376:379-384; Bromme, D. et al., 1996, J Biol Chem 271:2126-2132; Drake, F. H. et al., 1996, J Biol Chem 271:12511-12516, which are hereby incorporated by reference in their entirety. Concurrent to the cloning, the autosomal recessive disorder, pycnodysostosis, characterized by an osteopetrotic phenotype with a decrease in bone resorption, was mapped to mutations present in the cathepsin K gene. To date, all mutations identified in the cathepsin K gene are known to result in inactive protein. See Gelb, B. D. et al., 1996, Science 273:1236-1238; Johnson, M. R. et al., 1996, Genotie Res 6:1050-1055, which are hereby incorporated by reference in their entirety. Therefore, it appears that cathepsin K is involved in osteoclast mediated bone resorption.
Cathepsin K is synthesized as a 37 kDa pre-pro enzyme, which is localized to the lysosomal compartment and where it is presumably autoactivated to the mature 27 kDa enzyme at low pH. See McQueney, M. S. et al., 1997, J Biol Chem 272:13955-13960; Littlewood-Evans, A. et al., 1997, Bone 20:81-86, which are hereby incorporated by reference in their entirety. Cathepsin K is most closely related to cathepsin S having 56% sequence identity at the amino acid level. The S2P2 substrate specificity of cathepsin K is similar to that of cathepsin S with a preference in the P1 and P2 positions for a positively charged residue such as arginine, and a hydrophobic residue such as phenylalanine or leucine, respectively. See Bromme, D. et al., 1996, J Biol Chem 271: 2126-2132; Bossard, M. J. et al., 1996, J Biol Chem 271:12517-12524, which are hereby incorporated by reference in their entirety. Cathepsin K is active at a broad pH range with significant activity between pH 4-8, thus allowing for good catalytic activity in the resorption lacunae of osteoclasts where the pH is about 4-5.
Human type I collagen, the major collagen in bone is a good substrate for cathepsin K. See Kafienah, W., et al., 1998, Biochem J 331:727-732, which is hereby incorporated by reference in its entirety. In vitro experiments using antisense oligonucleotides to cathepsin K, have shown diminished bone resorption in vitro, which is probably due to a reduction in translation of cathepsin K mRNA. See Inui, T., et al., 1997, J Biol Chem 272:8109-8112, which is hereby incorporated by reference in its entirety. The crystal structure of cathepsin K has been resolved. See McGrath, M. E., et al., 1997, Nat Struct Biol 4:105-109; Zhao, B., et al., 1997, Nat Struct Biol 4: 109-11, which are hereby incorporated by reference in their entirety. Also, selective peptide based inhibitors of cathepsin K have been developed See Bromme, D., et al., 1996, Biochem J 315:85-89; Thompson, S. K., et al., 1997, Proc Natl Acad Sci USA 94:14249-14254, which are hereby incorporated by reference in their entirety. Accordingly, inhibitors of Cathepsin K can reduce bone resorption. Such inhibitors would be useful in treating disorders involving bone resorption, such as osteoporosis.
Compounds of the instant invention are useful as inhibitors of cathepsins. More particularly, the compounds of the instant invention are useful as inhibitors of Cathepsins K and L.
It is therefore an object of the invention to provide compounds which inhibit cathepsin activity in a mammal in need thereof.
It is another object of the invention to provide compounds which are useful for treating and/or preventing bone loss in a mammal in need thereof.
It is another object of the invention to provide compounds which are useful to reduce bone loss in a mammal in need thereof.
It is another object of the invention to provide compounds which are useful for treating and/or preventing bone fractures in a mammal in need thereof.
It is another object of the invention to provide compounds which are useful for treating and/or preventing osteoporosis in a mammal in need thereof.
It is another object of the invention to provide compounds which are useful for treating and/or preventing cathepsin dependent conditions or disease states in a mammal in need thereof.
The present invention relates to compounds of the following chemical formula: 
wherein
R1 and R2 are each independently selected from the group consisting of hydrogen, alkyl, oxo, xe2x80x94(CH2)pxe2x80x94NHxe2x80x94S(O)2xe2x80x94R3, xe2x80x94(CH2)pxe2x80x94NHxe2x80x94COxe2x80x94R4, xe2x80x94C(O)2R6, xe2x80x94(CH2)pOR5, xe2x80x94OR6, xe2x80x94(CH2)pNR7R8, xe2x80x94CN, xe2x80x94NH(CH2)pR3, xe2x80x94(CH2)pR3, xe2x80x94R3, xe2x80x94C(O)NHR6 and xe2x80x94C(O)NR6; or R1 and R2 can be joined together to form a system selected from the group consisting of aryl, cycloalkyl and heterocycloalkyl;
R3 is selected from the group consisting of aryl, arylalkyl, cycloalkyl, and heterocycloalkyl, wherein said aryl, arylalkyl and cycloalkyl groups are either unsubstituted or substituted with 1, 2 or 3 halogen atoms;
R4 is selected from the group consisting of aryl, cycloalkyl, heterocycloalkyl, biaryl, CH(R10)xe2x80x94NHC(O)2R3, OR5, (CH2)pR9, (CH2)p(R9)q, wherein said aryl, cycloalkyl, heterocycloalkyl and biaryl, groups are either unsubstituted or substituted with 1, 2 or 3 halogen atoms;
R5 is selected from the group consisting of alkyl, alkenyl, alkynyl, and (CH2)pR9;
R6 is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, and CH(R10)xe2x80x94NHC(O)2R3;
R7 and R8 are each independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, and xe2x80x94(CH2)pR3; or R7 and R8 are joined together to form a system selected from the group consisting of aryl and heterocycloalkyl;
R9 is selected from the group consisting of aryl, cycloalkyl and heterocycloalkyl;
R10 is selected from the group consisting of the side chains of the naturally occurring amino acids or unnaturally occurring amino acids;
each n is independently an integer from zero to four;
each p is independently an integer from zero to six;
each q is independently an integer from zero to four;
and the pharmaceutically acceptable salts thereof.
The present invention also relate to pharmaceutical compositions comprising the compounds of the present invention and a pharmaceutically acceptable carrier.
The present invention also relates to methods for making the pharmaceutical compositions of the present invention.
The present invention also relates to methods of inhibiting cathepsin activity and/or treating cathepsin dependent conditions in a mammal in need thereof comprising administering to the mammal the compounds and pharmaceutical compositions of the present invention.
The present invention also relates to methods of treating, preventing and/or reducing bone loss in a mammal in need thereof comprising administering to the mammal the compounds and pharmaceutical compositions of the present invention.
The present invention also relates to methods of inhibiting treating and/or preventing osteoporosis in a mammal in need thereof comprising administering to the mammal the compounds and pharmaceutical compositions of the present invention.
The present invention also relates to methods of reducing bone loss in a mammal in need thereof comprising administering to the mammal the compounds and pharmaceutical compositions of the present invention.
The present invention also relates to methods of treating and/or preventing bone fractures in a mammal in need thereof comprising administering to the mammal the compounds and pharmaceutical compositions of the present invention.
The present invention relates to the use of the compounds and pharmaceutical compositions of the present invention for the preparation of a medicament for treating or preventing bone loss in a mammal in need thereof.
The present invention relates to pharmaceutical compositions useful for treating or preventing bone loss in a mammal comprising a pharmaceutically effective amount of compounds of the present invention in association with pharmaceutically acceptable carriers.